CN107746840A - A kind of PA ase preparation and process for fixation - Google Patents
A kind of PA ase preparation and process for fixation Download PDFInfo
- Publication number
- CN107746840A CN107746840A CN201711208186.XA CN201711208186A CN107746840A CN 107746840 A CN107746840 A CN 107746840A CN 201711208186 A CN201711208186 A CN 201711208186A CN 107746840 A CN107746840 A CN 107746840A
- Authority
- CN
- China
- Prior art keywords
- ase
- cellulose nanocrystal
- fixation
- aqueous solution
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
- C12N11/12—Cellulose or derivatives thereof
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
- C12N9/84—Penicillin amidase (3.5.1.11)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y305/00—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5)
- C12Y305/01—Hydrolases acting on carbon-nitrogen bonds, other than peptide bonds (3.5) in linear amides (3.5.1)
- C12Y305/01011—Penicillin amidase (3.5.1.11), i.e. penicillin-amidohydrolase
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Mycology (AREA)
- Medicinal Chemistry (AREA)
- Immobilizing And Processing Of Enzymes And Microorganisms (AREA)
- Chemical Or Physical Treatment Of Fibers (AREA)
Abstract
The invention belongs to the technical field of material, discloses a kind of PA ase preparation and process for fixation.The process for fixation is：(1) under conditions of the gentle agitation of room, potassium hydroxide aqueous solution is added dropwise in the THPC aqueous solution, obtains the mixed liquor containing trihydroxy methyl phosphorus；(2) magnetic cellulose nanocrystal is added in the mixed liquor containing trihydroxy methyl phosphorus of step (1), 1 30min, centrifuge washing, the magnetic cellulose nanocrystal after being handled is stirred at room temperature；(3) PA ase is diluted in the cushioning liquid that pH is 6 11, obtains enzyme solutions；(4) magnetic cellulose nanocrystal after processing is added in enzyme solutions, cold curing, being fixed PA ase is PA ase preparation.Method of the invention is simple, cost is low；The prepared immobilized penicillin acylated enzyme enzyme activity rate of recovery is high, and the load factor of enzyme is high, while has preferable stability.
Description
Technical field
The invention belongs to the technical field of material, it is related to the fixing means and enzyme preparation of a kind of PA ase.
Background technology
Cellulose nanocrystal (CNC) is a kind of natural nano-material with premium properties, and it is cellulosic material one
The rhabdolith that the length obtained under fixed condition by sour water solution is 50-1000nm, width is 5-30nm.Cellulose nanocrystal exists
It is extremely stable in solution, it is difficult to separate.The magnetic cellulose that magnetic ferroferric oxide introducing Cellulose nanocrystal is prepared to obtain
It is nanocrystalline can be with liquid phase quick separating under magnetic fields.
Magnetic cellulose nanocrystal has high-crystallinity, high polymerization degree, high-hydrophilic, high mechanical properties, high bio-compatible
Property, high-specific surface area, the advantages that being easily recycled, there are the very big potentiality as enzyme immobilizatio carrier.
What magnetic cellulose nanocrystal immobilised enzymes was conventional at present is to be made using glutaraldehyde as crosslinking agent using cross-linking method
Enzyme is fixed on material surface.But glutaraldehyde has a significant impact to the activity of enzyme, the immobilised enzymes obtained using glutaraldehyde cross-linking
The enzyme activity rate of recovery is relatively low, at the same glutaraldehyde and also with carrier surface amino formed C=N key facile hydrolysis, particularly temperature compared with
Gao Shi, also make enzyme activity loss in immobilization process serious.
The content of the invention
The defects of in order to overcome prior art, primary and foremost purpose of the invention are to provide a kind of PA ase preparation
Process for fixation.For the present invention using trihydroxy methyl phosphorus as crosslinking agent, magnetic cellulose nanocrystal is carrier, and PA ase is entered
Preferable fixation is gone, the immobilised enzymes enzyme activity rate of recovery is high, and the load factor of enzyme is high, while has preferable stability.
Another object of the present invention is to provide the PA ase preparation being prepared by above-mentioned process for fixation.
The purpose of the present invention is achieved through the following technical solutions：
A kind of process for fixation of PA ase preparation, is comprised the steps of：
(1) under conditions of the gentle agitation of room, potassium hydroxide aqueous solution is added dropwise in the THPC aqueous solution,
Obtain the mixed liquor containing trihydroxy methyl phosphorus；
(2) magnetic cellulose nanocrystal is added in the mixed liquor containing trihydroxy methyl phosphorus of step (1), be stirred at room temperature
1-30min, centrifuge washing, the magnetic cellulose nanocrystal after being handled；
(3) PA ase is diluted in the cushioning liquid that pH is 6-11, obtains enzyme solutions；
(4) magnetic cellulose nanocrystal after processing is added in enzyme solutions, cold curing, being fixed penicillin acyl
It is PA ase preparation to change enzyme.
The concentration of the THPC aqueous solution described in step (1) is preferably 2.00g/90ml water -7.00g/90ml
Water；The concentration of potassium hydroxide aqueous solution described in step (1) is preferably 0.56g/10ml water -1.96g/10ml water；In step (1)
The volume ratio of the THPC aqueous solution reclaimed water and potassium hydroxide aqueous solution reclaimed water is 90:10.
It is preferably 8 minutes that the time is stirred at room temperature described in step (2)；The rotating speed being stirred at room temperature is 100~300rpm；
Cushioning liquid is preferably phosphate buffer solution described in step (3).
PH of buffer described in step (3) is preferably 7.5.
Described in step (4) in enzyme solutions PA ase with processing after magnetic cellulose nanocrystal in magnetic fibre
The nanocrystalline mass ratio of element is (2-7)：20(mg/mg).
The time of cold curing described in step (4) is 1h-12h.The cold curing is carried out under conditions of stirring.
Magnetic cellulose nanocrystal described in step (2) is the magnetic cellulose nanocrystal containing amino.
The specific preparation method of the magnetic cellulose nanocrystal is：
(S1) preparation of Cellulose nanocrystal body：Cellulosic material is added in acid solution, at 30-105 DEG C, stirring reaction
(0.5-2) h, after question response terminates, deacidified with distillation water washing, obtain Cellulose nanocrystal body；The concentration of the acid solution is
(1-14)mol/L；The solid-to-liquid ratio of the cellulosic material and acid solution is 1g:(5-100)ml；The acid solution is sulfuric acid, salt
One or more in acid, phosphoric acid, formic acid, acetic acid, citric acid and nitric acid；
(S2) Cellulose nanocrystal mixes complexing with molysite, chitosan：Calculated with iron ion molar concentration sum, by described in
Cellulose nanocrystal be added to concentration be (0.00005-1) mol/L iron salt solutions in, stir (0.1-3) h, then add
Mass concentration is 0.01-10% chitosan solution, stirs (0.1-3) h, obtains Cellulose nanocrystal-metal ion complex
Mixed liquor；The Cellulose nanocrystal and the solid-to-liquid ratio of iron salt solutions are 1g:(5-100)ml；The ferric iron accounts for total iron ion
Molal quantity is 50-66.7%；
(S3) it is crosslinked and precipitates：Adding mass concentration in the Cellulose nanocrystal-metal ion complex mixed liquor is
(0.01-10) % sodium tripolyphosphate solution, 1-60min is stirred, add aqueous slkali regulation pH to 8-13, be 20-90 in temperature
0.1-3h is reacted under the conditions of DEG C；Product is washed with deionized after reaction, then passes through and is dried to obtain magnetic fibre cellulosic material；Institute
The solid-to-liquid ratio for stating Cellulose nanocrystal and sodium tripolyphosphate solution is 1g:(1-500)ml.The cellulosic material is microcrystalline cellulose
Element, cellulose fibre, cotton, bamboo fibre, the one or more in flaxen fiber and bacteria cellulose.Step (S1), (S2) and
(S3) in, speed (1000-5000) r/min of the stirring.The aqueous slkali is NH3·H2O, NaOH, KOH or Ca (OH)2's
The aqueous solution.
The principle of the invention：
Potassium hydroxide aqueous solution is slowly dropped into i.e. available three hydroxyls of stirring in the THPC aqueous solution by the present invention
The methyl phosphorus aqueous solution.The present invention, as crosslinking agent, can be occurred using trihydroxy methyl phosphorus with carrier surface and the amido on enzyme surface
Mannich react, cross-linking reaction at room temperature can naturally-occurring, the P-CH formed2- NH keys are sufficiently stable, not facile hydrolysis,
It can preserve for a long time.The present invention PA ase is fixed in magnetic cellulose nanocrystal after, have wider array of temperature with
Scope applicable pH, is also easier to recycling.For the present invention from trihydroxy methyl phosphorus as crosslinking agent, the magnetic containing amino is fine
Dimension element is nanocrystalline to be used as carrier, and PA ase has been carried out into preferable fixation, and the immobilised enzymes enzyme activity rate of recovery is high, and enzyme is born
Load rate is high, while has preferable stability.The magnetic cellulose nanocrystal of the present invention has a high-specific surface area, high-crystallinity,
High polymerization degree, high-hydrophilic, high mechanical properties, good biocompatibility, it is easily recycled, is more beneficial for fixing penicillin acylation
Enzyme, improve the performance of immobilised enzymes.
The present invention is had the following advantages relative to prior art and effect：
(1) the trihydroxy methyl phosphorus that the present invention uses is prepared by THPC and potassium hydroxide, and price is just
Preferably, it is readily available, reduce the preparation cost for preparing immobilized penicillin acylated enzyme；
(2) P-CH formed between the trihydroxy methyl phosphorus and carrier and enzyme that the present invention uses2- NH keys are sufficiently stable, no
Facile hydrolysis, so the immobilized penicillin acylated enzyme being prepared is prepared relative to using conventional cross-linking agent glutaraldehyde cross-linking
PA ase there is more excellent zymologic property；
(3) the immobilised enzymes enzyme activity rate of recovery of the invention is high, and the load factor of enzyme is high, while has preferable stability.
Brief description of the drawings
The process that Fig. 1 prepares immobilized penicillin acylated enzyme for embodiment 1~6 using trihydroxy methyl phosphorus as crosslinking agent is shown
It is intended to；MNCC is the magnetic cellulose nanocrystal containing amino, PA-NH2For PA ase；
Fig. 2 is that immobilized penicillin acylated enzyme prepared by embodiment 1 and free PA ase are steady at different temperatures
Qualitative comparison diagram；
Fig. 3 is that immobilized penicillin acylated enzyme prepared by embodiment 1 is stablized from free PA ase under different pH
Property comparison diagram.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Magnetic cellulose nanocrystal is prepared by the following method to obtain in embodiment 1~6：Cellulose fibre is added
In 6mol/L formic acid solutions, wherein the solid-to-liquid ratio of microcrystalline cellulose and acid solution is 1:25g/mL, mixed liquor and 60 DEG C of reactions, are stirred
Speed 1000r/min is mixed, after question response terminates, reactant is moved into centrifuge tube, 5min is centrifuged under 4000r/min, goes
Supernatant, deionized water is added, wherein deionized water is identical with the volume for adding acid solution before, under 4000r/min
5min is centrifuged, obtains Cellulose nanocrystal crude product.6.6g Cellulose nanocrystal crude products are taken, add 200ml iron concentrations
For 0.3782mol/L solution, wherein Fe3+It is 60%, Fe to account for the ratio between total iron ion molal quantity2+40% is accounted for, is added after stirring 10min
Enter 2% chitosan solution 30ml, stir speed (S.S.) 1000r/min, stir 50min.Then add 20ml and contain 6% sodium tripolyphosphate
The aqueous solution stirs 1min, adds the ammoniacal liquor of 10ml 28%, reacts 40min at 80 DEG C.By products obtained therefrom distillation water washing, freezing
Magnetic cellulose nanocrystal is obtained after drying.
Embodiment 1~6 prepares the process schematic of immobilized penicillin acylated enzyme using trihydroxy methyl phosphorus as crosslinking agent
As shown in figure 1, MNCC is the magnetic cellulose nanocrystal containing amino, PA-NH2For PA ase.
Embodiment 1
(1) THPC for taking quality to be 2.00g is added in 90ml water, and it is water-soluble to obtain THPC
Liquid；The potassium hydroxide for taking quality to be 0.56g is added in 10ml water, obtains potassium hydroxide aqueous solution；
(2) at room temperature, potassium hydroxide aqueous solution is slowly dropped into the THPC aqueous solution, obtains mixing molten
Liquid A；
(3) 2.00g magnetic cellulose nanocrystals are added in mixed solution A, are stirred at room temperature that (rotating speed of stirring is
150rpm) 8min, centrifugation, is washed with deionized, the magnetic cellulose nanocrystal after being handled；
(4) take 200.00mg PA ases to be dissolved in the phosphate buffer that 50ml pH are 7.5, obtain enzyme solutions；
(5) enzyme solutions for being configured the magnetic cellulose nanocrystal after processing resulting in step (3) with step (4)
Mixing, (rotating speed of stirring is 150rpm) 3h, deionized water water washing, being fixed PA ase are stirred at room temperature；
The enzyme activity rate of recovery 79.2%, enzyme carrying capacity 76.8mg/g.
Immobilized penicillin acylated enzyme and free PA ase prepared by embodiment 1 stability pair at different temperatures
It is more as shown in Figure 2 than scheming；Immobilized penicillin acylated enzyme prepared by embodiment 1 stabilization under different pH from free PA ase
Property comparison diagram is as shown in Figure 3.
Embodiment 2
(1) THPC for taking quality to be 7.00g is added in 90ml water, and it is water-soluble to obtain THPC
Liquid；The potassium hydroxide for taking quality to be 1.96g is added in 10ml water, obtains potassium hydroxide aqueous solution；
(2) at room temperature, potassium hydroxide aqueous solution is slowly dropped into the THPC aqueous solution, obtains mixing molten
Liquid A；
(3) 2.00g magnetic cellulose nanocrystals are added in mixed solution A, are stirred at room temperature that (rotating speed of stirring is
150rpm) 8min, centrifugation, is washed with deionized, the magnetic cellulose nanocrystal after being handled；
(4) take 200.00mg PA ases to be dissolved in the phosphate buffer that 50ml pH are 7.5, obtain enzyme solutions；
(5) it is the magnetic cellulose nanocrystal after the processing obtained by step (3) and enzyme that step (4) is configured is molten
Liquid mixes, and stirs (rotating speed of stirring is 150rpm) 3h, being fixed PA ase after washing, enzyme activity recovery at room temperature
Rate 41.4%, enzyme load capacity 145.5mg/g.
Embodiment 3
(1) THPC for taking quality to be 2.00g is added in 90ml water, and it is water-soluble to obtain THPC
Liquid；The potassium hydroxide for taking quality to be 0.56g is added in 10ml water, obtains potassium hydroxide aqueous solution；
(2) at room temperature, potassium hydroxide aqueous solution is slowly added dropwise in the THPC aqueous solution, mixed
Close solution A；
(3) 2.00g magnetic cellulose nanocrystals are added in mixed solution A, (150rpm) 8min is stirred at room temperature, from
The heart, it is washed with deionized, the magnetic cellulose nanocrystal after being handled；
(4) take 600.00mg PA ases to be dissolved in the phosphate buffer that 50ml pH are 7.5, obtain enzyme solutions；
(5) it is the magnetic cellulose nanocrystal after the processing obtained by step (3) and enzyme that step (4) is configured is molten
Liquid mixes, and stirs (rotating speed of stirring is 150rpm) 3h, being fixed PA ase after washing at room temperature；Enzyme activity reclaims
Rate 67.5%, enzyme load capacity 170.5mg/g.
Embodiment 4
(1) THPC for taking quality to be 5.00g is added in 90ml water, and it is water-soluble to obtain THPC
Liquid；The potassium hydroxide for taking quality to be 1.40g is added in 10ml water, obtains potassium hydroxide aqueous solution；
(2) at room temperature, potassium hydroxide aqueous solution is slowly added dropwise in the THPC aqueous solution, mixed
Close solution A；
(3) 2.00g magnetic cellulose nanocrystals are added in mixed solution A, after (150rpm) 8min is stirred at room temperature, from
The heart, it is washed with deionized, the magnetic cellulose nanocrystal after being handled；
(4) take 200.00mg PA ases to be dissolved in the phosphate buffer that 50ml pH are 7.5, obtain enzyme solutions；
(5) enzyme solutions for being configured the magnetic cellulose nanocrystal after processing resulting in step (3) with step (4)
Mixing, (150rpm) 3h, being fixed PA ase after deionized water washing are stirred at room temperature；The enzyme activity rate of recovery
67.3%, enzyme load capacity 135.0mg/g.
Embodiment 5
(1) THPC for taking quality to be 3.00g is added in 90ml water, and it is water-soluble to obtain THPC
Liquid；The potassium hydroxide for taking quality to be 0.84g is added in 10ml water, obtains potassium hydroxide aqueous solution；
(2) at room temperature, potassium hydroxide aqueous solution is slowly dropped into the THPC aqueous solution, obtains mixing molten
Liquid A；
(3) 2.00g magnetic cellulose nanocrystals are added in mixed solution A, (150rpm) 8min is stirred at room temperature, from
The heart, it is washed with deionized, the magnetic cellulose nanocrystal after being handled；
(4) take 600.00mg PA ases to be dissolved in the phosphate buffer that 50ml pH are 7.5, obtain enzyme solutions；
(5) enzyme solutions for being configured the magnetic cellulose nanocrystal after processing resulting in step (3) with step (4)
Mixing, (150rpm) 10h, being fixed PA ase after deionized water washing are stirred at room temperature；The enzyme activity rate of recovery
71.3%, enzyme load capacity 127.8mg/g.
Embodiment 6
(1) THPC for taking quality to be 4.00g is added in 90ml water, and it is water-soluble to obtain THPC
Liquid；The potassium hydroxide for taking quality to be 1.12g is added in 10ml water, obtains potassium hydroxide aqueous solution；
(2) at room temperature, potassium hydroxide aqueous solution is slowly dropped into the THPC aqueous solution, obtains mixing molten
Liquid A；
(3) 2.00g magnetic cellulose nanocrystals are added in mixed solution A, 8min are stirred at room temperature, centrifuge, spend from
Sub- water washing, the magnetic cellulose nanocrystal after being handled；
(4) take 600.00mg PA ases to be dissolved in the phosphate buffer that 50ml pH are 7.5, obtain enzyme solutions；
(5) enzyme solutions for being configured the magnetic cellulose nanocrystal after processing resulting in step (3) with step (4)
Mixing, (150rpm) 9h, being fixed PA ase after deionized water washing are stirred at room temperature；The enzyme activity rate of recovery
67.2%, enzyme load capacity 177.3mg/g.
Above-described embodiment is only the section Example of the present invention, is not used for limiting the practical range of the present invention；It is i.e. all according to
The equivalent changes and modifications that present invention is made, all covered by the claims in the present invention scope claimed.
Claims (7)
- A kind of 1. process for fixation of PA ase preparation, it is characterised in that：Comprise the steps of：(1) under conditions of the gentle agitation of room, potassium hydroxide aqueous solution is added dropwise in the THPC aqueous solution, obtained Mixed liquor containing trihydroxy methyl phosphorus；(2) magnetic cellulose nanocrystal is added in the mixed liquor containing trihydroxy methyl phosphorus of step (1), 1- is stirred at room temperature 30min, centrifuge washing, the magnetic cellulose nanocrystal after being handled；(3) PA ase is diluted in the cushioning liquid that pH is 6-11, obtains enzyme solutions；(4) magnetic cellulose nanocrystal after processing is added in enzyme solutions, cold curing, being fixed PA ase That is PA ase preparation.
- 2. the process for fixation of PA ase preparation according to claim 1, it is characterised in that：Described in step (1) The concentration of the THPC aqueous solution is 2.00g/90ml water -7.00g/90ml water；Potassium hydroxide water described in step (1) The concentration of solution is 0.56g/10ml water -1.96g/10ml water；THPC aqueous solution reclaimed water described in step (1) with The volume ratio of potassium hydroxide aqueous solution reclaimed water is 90:10.
- 3. the process for fixation of PA ase preparation according to claim 1, it is characterised in that：Described in step (2) Magnetic cellulose nanocrystal is the magnetic cellulose nanocrystal containing amino.
- 4. the process for fixation of PA ase preparation according to claim 1, it is characterised in that：Described in step (3) Cushioning liquid is phosphate buffer solution.
- 5. the process for fixation of PA ase preparation according to claim 1, it is characterised in that：Described in step (3) PH of buffer be 7.5.
- 6. the process for fixation of PA ase preparation according to claim 1, it is characterised in that：Described in step (4) PA ase and the mass ratio of magnetic cellulose nanocrystal in the magnetic cellulose nanocrystal after processing are (2- in enzyme solutions 7)：20.
- 7. the process for fixation of PA ase preparation according to claim 1, it is characterised in that：Described in step (4) The time of cold curing is 1h-12h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711208186.XA CN107746840B (en) | 2017-11-27 | 2017-11-27 | Penicillin acylase preparation and immobilization method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711208186.XA CN107746840B (en) | 2017-11-27 | 2017-11-27 | Penicillin acylase preparation and immobilization method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107746840A true CN107746840A (en) | 2018-03-02 |
| CN107746840B CN107746840B (en) | 2020-12-22 |
Family
ID=61252472
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711208186.XA Active CN107746840B (en) | 2017-11-27 | 2017-11-27 | Penicillin acylase preparation and immobilization method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107746840B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109207466A (en) * | 2018-11-08 | 2019-01-15 | 山东鲁抗医药股份有限公司 | A kind of process for fixation and immobilised enzymes of PA ase |
| CN110846306A (en) * | 2019-12-09 | 2020-02-28 | 江苏省中国科学院植物研究所 | Amphiphilic enzyme immobilization carrier |
| CN113462680A (en) * | 2021-07-28 | 2021-10-01 | 兰州理工大学 | Preparation of magnetic immobilized penicillin G acylase doped with divalent manganese ion |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1618959A (en) * | 2004-10-12 | 2005-05-25 | 上海大学 | Alpha-glucosidase immobilization method |
| CN103709422A (en) * | 2013-12-20 | 2014-04-09 | 华南理工大学 | Preparation method of magnetic cellulose nanocrystal |
-
2017
- 2017-11-27 CN CN201711208186.XA patent/CN107746840B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1618959A (en) * | 2004-10-12 | 2005-05-25 | 上海大学 | Alpha-glucosidase immobilization method |
| CN103709422A (en) * | 2013-12-20 | 2014-04-09 | 华南理工大学 | Preparation method of magnetic cellulose nanocrystal |
Non-Patent Citations (4)
| Title |
|---|
| BAHULEKAR, RV等: "Immobilization of penicillin G acylase on functionalized macroporous polymer beads", 《POLYMER》 * |
| 乌云高娃等: "青霉素酰化酶在甲基丙烯酸缩水甘油酯共聚物上的固定化", 《催化学报》 * |
| 李斌等: "《食品酶学与酶工程》", 30 September 2017, 中国农业大学出版社 * |
| 江连洲等: "《酶在大豆制品中的应用》", 31 August 2015, 中国轻工业出版社 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109207466A (en) * | 2018-11-08 | 2019-01-15 | 山东鲁抗医药股份有限公司 | A kind of process for fixation and immobilised enzymes of PA ase |
| CN109207466B (en) * | 2018-11-08 | 2021-01-08 | 山东鲁抗医药股份有限公司 | Immobilization method of penicillin acylase and immobilized enzyme |
| CN110846306A (en) * | 2019-12-09 | 2020-02-28 | 江苏省中国科学院植物研究所 | Amphiphilic enzyme immobilization carrier |
| CN110846306B (en) * | 2019-12-09 | 2021-08-17 | 江苏省中国科学院植物研究所 | Amphiphilic enzyme immobilization carrier |
| CN113462680A (en) * | 2021-07-28 | 2021-10-01 | 兰州理工大学 | Preparation of magnetic immobilized penicillin G acylase doped with divalent manganese ion |
| CN113462680B (en) * | 2021-07-28 | 2023-08-22 | 兰州理工大学 | Preparation of Magnetically Immobilized Penicillin G Acylase Doped with Divalent Manganese Ions |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107746840B (en) | 2020-12-22 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107746840A (en) | A kind of PA ase preparation and process for fixation | |
| CN103709422B (en) | A kind of preparation method of magnetic cellulose nanocrystal | |
| Cao et al. | Cellulose hydrolysis using zinc chloride as a solvent and catalyst | |
| CN110600735B (en) | Method for preparing lithium iron phosphate cathode material at low cost and application | |
| Huacong et al. | Immobilization of penicillin G acylase on magnetic nanoparticles modified by ionic liquids | |
| Wan et al. | α-glucosidase immobilization on magnetic core-shell metal-organic frameworks for inhibitor screening from traditional Chinese medicines | |
| CN105524909A (en) | Magnetic chitosan microsphere for enzyme immobilization as well as preparation method and application thereof | |
| KR101347205B1 (en) | Enzyme immobilized on Au-doped Magnetic silica nanoparticle, method for producing the same, and hydrolytic degradation method of biomass thereby | |
| CN101462942A (en) | Method for producing high purity battery level iron oxalate from pickle liquor | |
| CN106011206A (en) | Method for preparing bioactive peptide from immobilized bi-enzyme of composite carrier magnetic nano-particles | |
| CN102517276B (en) | Method for preparing magnetic nano carrier immobilized aldolase with high substrate tolerance | |
| CN105695442A (en) | Modified magnetic chitosan microspheres for enzyme immobilization and preparation method and application thereof | |
| CN106480130B (en) | A kind of method that magnetic Nano immobilized enzyme hydrolyzing straw can be recycled in application | |
| CN113462680B (en) | Preparation of Magnetically Immobilized Penicillin G Acylase Doped with Divalent Manganese Ions | |
| CN114984926A (en) | Preparation method of high-agglomeration-resistance RGO-based magnetic lithium ion imprinted polymer | |
| CN111573680A (en) | Method for removing iron in quartz sand | |
| CN113073095A (en) | High-enzyme-activity magnetic immobilized laccase and preparation method thereof, and method for efficiently degrading dye | |
| WO2022062836A1 (en) | Pyrrolic nitrogen-dominated nitrogen-doped graphene synthesis method and pyrrolic nitrogen-dominated nitrogen-doped graphene prepared by method | |
| CN113817179A (en) | Preparation method of nanofiber magnetofluid, nanofiber magnetofluid and application | |
| CN109321559B (en) | Magnetic Fe3O4Method for immobilizing fructosyltransferase by taking polysaccharide microspheres as carrier | |
| CN113136381B (en) | Preparation method of multienzyme conjoined nano reactor and application thereof in synchronous synthesis | |
| CN101987737A (en) | Method for preparing spectrum pure calcium carbonate | |
| CN111592449A (en) | Production process of benzoin | |
| CN110551225A (en) | preparation method of dextrin with thermal viscosity stability | |
| CN116589609B (en) | Preparation method of phosphorylated nano-chitin based on mechanochemical method and phosphorylated nano-chitin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |